PROJECT SUMMARY ? ANALYTICAL IMAGING FACILITY SHARED RESOURCE The Analytical Imaging Facility (AIF) is a comprehensive microscopy shared resource that offers a broad spectrum of imaging technologies to AECC investigators. These include: macro-imaging with a stereoscope, classical microscopy including brightfield, darkfield, phase contrast, Nomarski differential interference contrast and wide-field epifluorescence, confocal microscopy for cell culture and tissues, high-speed and high-capacity automatic slide scanning, high speed spinning disk confocal microscopy for live cells, intravital multiphoton microscopy for live animals, super-resolution STORM and SIM microscopy, standard transmission and scanning electron microscopy (TEM & SEM), cryoEM of macromolecules, electron tomography and 3D reconstruction of high pressure frozen cells and tissue, automated imaging of serial sections by field emission SEM (FESEM) and SEM of frozen hydrated cells and tissues. The wide range of imaging modalities is especially important for studying the diverse biological models utilized by the AECC research community. For instance, cells labeled with fluorescent reporter genes may be imaged at the whole animal or whole organ level down to subcellular and molecular levels. Over the span of 23 years, the AIF has expanded its dedicated laboratory space and staff, and added multiple new instruments and equipment upgrades. This growth has been enabled by funding from Cancer Center and College and Shared Instruments Grants. This facility furnishes AECC investigators with the majority of their on-site access to routine and sophisticated imaging technologies. The technologies provided are essential to the research of many AECC members, and cannot be outsourced or obtained at other sites in any practical or cost effective way. Having the technology and staff in one location provides continuity, facilitates cross-training and adds value to the AECC researchers who use microscopy. Beyond making routine and complex imaging technologies available to the AECC community, the AIF also provides comprehensive training in most technologies so that AECC members and their staff are competent in the operations of the equipment, image analysis and the interpretation of the data acquired. This is accomplished by supporting routine as well as advanced microscopy applications, training users in microscopy and image analysis software, hosting regular workshops on microscopy and image processing and introducing innovative imaging technologies developed at this center to meet the needs of AECC investigators.